Effective targeting of prostate cancer by lymphocytes redirected by a PSMA×CD3 bispecific single-chain diabody

Kerstin Fortmüller, Karen Alt, Dorothee Gierschner, Philipp Wolf, Volker Baum, Nikolaus Freudenberg, Ulrich Wetterauer, Ursula Elsässer-Beile, Patrick Bühler

Research output: Contribution to journalArticleResearchpeer-review

30 Citations (Scopus)

Abstract

BACKGROUND For redirecting T-lymphocytes to induce prostate cancer cell lysis, we constructed a novel bispecific single-chain (bsc) diabody directed to the prostate specific membrane antigen (PSMA) and the T-cell receptor (TCR)-associated CD3 molecule on T-cells. METHODS The PSMA×CD3 bsc diabody was generated from an anti-CD3 single chain Fv fragment (scFv) and the anti-PSMA scFv D7. It was expressed in E. coli and purified from the periplasmic extract and culture supernatant by immobilized metal affinity chromatography (IMAC). The binding properties were tested on PSMA-expressing prostate cancer cells and PSMA-negative cell lines as well as on Jurkat cells by flow cytometry. For in vitro functional analysis, a cell viability test (WST-1) was used and activation of T-cells was determined by measuring the surface marker expression of CD25 and CD69. For in vivo evaluation, the diabody was administered in combination with human peripheral blood lymphocytes (Ly) in a C4-2 xenograft-SCID mouse model. RESULTS Specific binding of the PSMA×CD3 bsc diabody both to CD3-positive Jurkat cells and PSMA-expressing C4-2 cells was shown by flow cytometry. In vitro, the PSMA×CD3 bsc diabody proved to be a potent agent for retargeting CD4+ and CD8+ human lymphocytes to lyse C4-2 prostate cancer cells. Treatment of SCID mice bearing C4-2 tumor xenografts with the diabody and human lymphocytes efficiently inhibited tumor growth. CONCLUSIONS The PSMA×CD3 bsc diabody bears a high potential for the immunotherapy of prostate cancer.

Original languageEnglish
Pages (from-to)588-596
Number of pages9
JournalThe Prostate
Volume71
Issue number6
DOIs
Publication statusPublished - May 2011
Externally publishedYes

Keywords

  • bispecific diabody
  • immunotherapy
  • prostate cancer
  • PSMA×CD3 bsc diabody
  • T-cell redirecting

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